Aim: This study aims to conduct a comparative evaluation of the impact of sodium hypochlorite and strawberry extract as irrigation solutions through the utilization of cone focal laser scanning microscopy. Methods: preparation and sterilization of single-rooted premolars (n = 21).A mature biofilm of Enterococcus faecalis was cultivated.The roots were separated into three groups based on the activation technique employed, namely ultrasonic, sonic, and positive control. The study involved the division of each group into two subgroups (n=7) based on the type of irrigant utilized:strawberry extract and 2.5% sodium hypochlorite. The specimens were prepared by cutting and subsequently analyzed by confocal laser scanning microscopy. The fluorescent images were subjected to analysis using Zen imaging software. The data analysis was conducted utilizing the one-way analysis of variance (ANOVA) method, followed by Tukey's honestly significant difference (HSD) test for pairwise comparisons. The level of statistical significance was established at a significance level of 5%. Results: The Ultrasonic and Sonic stimulation groups exhibited a greater proportion of deceased cells in comparison to the positive control group for both the NaOCl and Strawberry Extract groups. In the NaOCL group,the ultrasonic activation demonstrated the highest proportion of deceased cells, followed by the sonic activation and the positive control without activation, with percentages of deceased cells amounting to 57.229%± 3.998, 47.086%± 4.558, and 29.457%± 3.713, respectively.it was concluded that Strawberry extract has been shown to exhibit more bacterial reduction with Ultrasonic activation than with Sonic or non-activation approaches, making it a viable irrigation alternative for endodontic therapy.